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Aurora B kinase inhibitor 3 - hydroxyflavone anti-tumor effect of the cum Ly-6 gene family members LYPD6 cloning and characterization analysis

Author: ZhangZuoFeng
Tutor: YuLong
School: Fudan University
Course: Genetics
Keywords: Aurora Gene family LYPD6 Ly-6 Flavonoids Hydroxy Anti-tumor effects Kinase inhibitors Functional studies cDNA Transcriptional activity Domain Human testis Transfection Library Subcellular localization Activity Detection Kinase activity Tumor cells Gene
CLC: Q343
Type: Master's thesis
Year: 2010
Downloads: 18
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Abstract


This thesis consists of two parts of the study work . The first part of the analysis 3 - hydroxyflavone internal suppression of endogenous Aurora B kinase activity and anti-tumor effect , the second part on human LYPD6 gene cloning and functional studies . Through these two studies , we found that : (1) high-throughput screening , get a new Aurora B kinase inhibitor - 3 - hydroxyflavone . In vitro activity tests proved , 3 - hydroxyflavone inhibiting Aurora B IC50 in nmol magnitude . Cytology experimental results show that 3 - hydroxyflavone can greatly reduce the phosphorylation of histone H3 , indicating that the inhibitory activity of endogenous Aurora B . The BIAcore detect 3 - hydroxyflavone and Aurora B protein interactions , and the results show that both the dissociation constant of 1.01 × 10- mol / L . 3 - the hydroxyflavone In 5μM concentration can inhibit tumor cell colony formation rate and reduce cell viability , is also reflected in the administration period of tumor cell growth inhibition , withdrawal and then resumed rapid growth proved 3 - hydroxyflavone inhibit tumor cell the rapid proliferation . (2) cloned from a human testis cDNA library in a new LU domain new gene LYPD6 of LU protein structural domain of about 80 amino acids , with 10 conserved cysteine ??residues in the formation of particular disulfide bond structure . Therefore, this new gene defined as a new member of the Ly-6 gene family , which is located on chromosome 2q23.1 - 23.2 . LYPD6 cDNA length of 3501bp , contains an open reading frame encoding 171 amino acids . The subcellular localization of over-expression in COS-7 cells , LYPD6 localized in the cytoplasm . RT-PCR showed LYPD6 widely present in the human tissue , but the mind , higher expression levels in the brain . In addition, overexpression LYPD6 in HEK-293T cells can inhibit the transcriptional activity of the AP1 pathway . The above two parts , respectively, proved that 3 - hydroxyflavone inhibit the transcriptional activity of the rapid proliferation of tumor cells as well as in HEK-293T cells the overexpression LYPD6 can be suppressed AP1 pathway . Laid a solid foundation for further study in the future .

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CLC: > Biological Sciences > Genetics > Genetics subdiscipline > Cytogenetic
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